Isolation of Amniotic Fluid Mesenchymal Stem Cells Obtained from Cesarean Sections
DOI:
https://doi.org/10.3889/oamjms.2020.3848Keywords:
Amniotic fluid, Mesenchymal stem cells, Cesarean sectionAbstract
BACKGROUND: The term amniotic fluid (AF) can be an ideal alternative as a source of mesenchymal stem cells (MSCs), originating from the neonate. Preclinical studies of the second- and third-trimester amnion fluid cells confirmed the number of potential donors from this wasted material.
AIM: This study aims to look at the forming time of stem cells derived from AF-MSCs.
MATERIALS AND METHODS: AF samples from healthy human donors were collected during full-term C-sections and kept at 4°C until processed. The number of colony-forming unit-fibroblast was assessed microscopically by calculating spindle-shaped colonies that clearly resembled of fibroblasts and did not include colonies with rounded epithelioid morphology. The immunophenotyping of their independent AF preparations was done using the human MSC phenotyping kit which was done according to the manufacturer’s instruction by flow cytometry.
RESULTS: The result showed that it succeeded in getting 8 million cells which will be used for research on pelvic organ prolapse therapy using AF-MSCs. The stem cell isolation totally takes 6 weeks. We got 2 million stem cells in one flask.
CONCLUSION: This study concludes that the time needed for differentiation of AF-MSCs is 6 weeks and AF-MSCs express mesenchymal markers such as CD90, CD73 (SH3, SH), and CD105 (SH2) and these cells also express HLA antigens – ABC, CD 34, and CD 45 which are hematopoietic markers and endothelial CD31 markers.
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DeSacco SD, DeFilippo RE, Perin L. Amniotic fluid as a source of pluripotent and multipotent stem cells for organ regeneration. Curr Opin Organ Transplant. 2011;16(1):101-5. https://doi. org/10.1097/mot.0b013e3283424f6e PMid:21157345
Roubelakis MG, Trohatou O, Anagnou NP. Amniotic fluid and amniotic membrane stem cells: Marker discovery. Stem Cells Int. 2012;2012:1-9. https://doi.org/10.1155/2012/107836
Asl KD, Shafei H, Rad JS, Nozad HO. Comparison of characteristics of human amniotic membrane and human adipose tissue derived mesenchymal stem cells. World J Plast Surg. 2016;6(1):33-9. PMid:28289611
Gaafar TM, Hawary RE, Osman A, Attia W, Hamza H, Brockmeier K. Comparative characteristics of amniotic membrane, endometrium and ovarian derived mesenchymal stem cells: A role for amniotic membrane in stem cell therapy. Middle East Fertil Soc J. 2014;19(3):156-70 https://doi. org/10.1016/j.mefs.2014.01.002
Moraghebi R, Kirkeby A, Chaves P, Rönn RE, Sitnicka E, Parmar M, et al. Term amniotic fluid: An unexploited reserve of mesenchymal stromal cells for reprogramming and potential cell therapy applications. Stem Cell Res Ther. 2017;8(190):1- 12. https://doi.org/10.1186/s13287-017-0582-6
Kim BS, Chun SY, Lee JK, Lim HJ, Bae JS, Chung HY, et al. Human amniotic fluid stem cell injection therapy for urethral sphincter regeneration in animal model. BMC Med. 2012;10:94. https://doi.org/10.1186/1741-7015-10-94 PMid:22906045
Decoppi P, Bartsch G Jr., Siddiqui MM, Xu T, Santos CC, Perin L, et al. Isolation of amniotic stem cell lines with potential for therapy. Nat Biotechnol. 2007;25(1):100-6. PMid:17206138
Savickiene J, Treigyte G, Baronaite S, Valiuliene G, Kaupinis A, Valius M. et al. Human amniotic fluid mesenchymal stem cells from second-and third-trimester amniocentesis: Differentiation potential, molecular signature, and proteome analysis. Stem Cells Int. 2015;2015:319238. https://doi.org/10.1155/2015/319238 PMid:26351462
Tsai MS, Lee JL, Chang YJ, Hwang SM. Isolation of human multipotent mesenchymal stem cells from second-trimester amniotic fluid using a novel two-stage culture protocol. Hum Reprod. 2004;19(6):1450-6. https://doi.org/10.1093/humrep/ deh279 PMid:15105397
Spitzhorn LS, Raman MS, Schwindt L, Ho HT, Wruck W, Bohndorf M, et al. Isolation and molecular characterization of amniotic fluid-derived mesenchymal stem cells obtained from caesarean sections. Stem Cells Int. 2017;2017:5932706. https://doi.org/10.1155/2017/5932706 PMid:29225627
Haasters F, Prall WC, Anz D, Bourquin C, Pautke C, Endres S, et al. Morphological and immunocytochemical characteristics indicate the yield of early progenitors and represent a quality control for human mesenchymal stem cell culturing. J Anat. 2009;214(5):759-67. https://doi. org/10.1111/j.1469-7580.2009.01065.x PMid:19438770
Kim EY, Lee KB, Kim MK. The potential of mesenchymal stem cells derived from amniotic membrane and amniotic fluid for neuronal regenerative therapy. BMB Rep. 2014;47(3):135-40. https://doi.org/10.5483/bmbrep.2014.47.3.289 PMid:24499672
Larson A, Gallichio VS. Amniotic derived stem cells: Role and function in regenerative medicine. J Cell Sci Ther. 2017;8(3):1-10.
Steigman SA, Fauza DO. Isolation of mesenchymal stem cells for amniotic fluid and placenta. Curr Protocol Stem Cell Biol. 2007;1:E2. https://doi.org/10.1002/9780470151808.sc01e02s1 PMid:18785167
Chamberlain G, Fox J, Ashton B, Middleton J. Concise review: Mesenchymal stem cells: Their phenotype, differentiation capacity, immunological features, and potential for homing. Stem Cells. 2007;25(11):2739-49. https://doi.org/10.1634/ stemcells.2007-0197 PMid:17656645
Lim R. Concise review: Fetal membranes in regenerative medicine: New tricks from an old dog? Stem Cells Transl Med. 2017;6(9):1767-76. https://doi.org/10.1002/sctm.16-0447 PMid:28834402
Dominici M, Blanc KL, Mueller I, Slaper-Cortenbach I, Marini F, Krause D, et al. Position paper: Minimal criteria for defining multipotent mesenchymal stromal cells. The international society for cellular therapy position statement. Cytotherapy. 2006;8(4):315-7. https://doi.org/10.1080/14653240600855905 PMid:16923606
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Copyright (c) 2020 Bobby Indra Utama, Afriwardi Afriwardi, Budi Iman Santoso, Hirowati Ali (Author)
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